Polymerase chain reaction: amplification of DNA from fixed tissue.
The polymerase chain reaction (PCR) allows the analysis of DNA from biologic samples containing only nanogram quantities of DNA. We used DNA purified from fresh or frozen peripheral blood (PB) leukocytes and formalin, or B-5 fixed bone marrow aspirate clots (BM). A sequence of the beta-globin gene was amplified via the PCR then hybridized with allele specific oligonucleotide probes for hemoglobin A, S, and C. All DNA preparations, including formalin and B-5 fixed BMs, were successfully amplified; the hybridization of the amplified products resulted in patterns consistent with the hemoglobin phenotype for all patients. PCR can be used on DNA from many sources; retrospective studies using paraffin embedded fixed tissue are possible because extremely small amounts of DNA present in fixed tissue can be successfully amplified.
Crisan
D
D
Department of Pathology, University of Pittsburgh, PA 15213-2582.
Cadoff
E M
EM
Mattson
J C
JC
Hartle
K A
KA
D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001483:Base Sequence; D004247:DNA / Q000235:genetics; D015342:DNA Probes; D004277:DNA, Single-Stranded / Q000235:genetics; D005260:Female; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D016133:Polymerase Chain Reaction / Q000379:methods*; D013048:Specimen Handling / Q000379:methods
D015342:DNA Probes; D004277:DNA, Single-Stranded; D005404:Fixatives; D005557:Formaldehyde; D004247:DNA